Achromobacter sp. D-12 and application thereof in microbial degradation of acetochlor

A technology for the degradation of Achromobacter and microorganisms, which is applied in the direction of microorganism-based methods, microorganisms, biological water/sewage treatment, etc., can solve the problems that Achromobacter has not been found to degrade acetochlor, etc., and achieves low cost and simple preparation process , the effect of good application prospects

Active Publication Date: 2013-04-10
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] Achromobacter in the present invention is a common short bacillus. After searching patents and

Method used

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  • Achromobacter sp. D-12 and application thereof in microbial degradation of acetochlor
  • Achromobacter sp. D-12 and application thereof in microbial degradation of acetochlor
  • Achromobacter sp. D-12 and application thereof in microbial degradation of acetochlor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1: Screening and identification of strains

[0042] 1) culture medium

[0043] Inorganic salt culture medium: NaCl 1g, K 2 HPO 4 1.5g, KH 2 PO 4 0.5g, (NH4) 2 SO 4 1.5g, MgSO 4 0.1g, 1ml trace element solution, made up to 1000ml with distilled water, mixed and stirred evenly, natural pH value, prepared after high-pressure steam sterilization (121°C, 20min), and each liter of trace element solution contains MnSO 4 ·H 2 O 0.13g, ZnCl 2 0.23g, CuSO 4 ·H 2 O 0.03g, CoCl 2 ·6H 2 O 0.42g, Na 2 MoO 4 2H 2 O 0.15g, AlCl 3 ·6H 2 O 0.05g, make up to 1000ml with distilled water.

[0044] Enrichment culture solution: Add acetochlor solution to the inorganic salt culture solution so that the concentration of acetochlor is 100 mg / L.

[0045] LB culture medium: Yeast powder 10g, peptone 5.0g, sodium chloride 10.0g, distilled water to make up to 1000ml, mixed and stirred evenly, natural pH value, prepared after high-pressure steam sterilization (121°C...

Embodiment 2

[0052] Embodiment 2: the preparation that contains bacterial cell suspension

[0053] (1) Slant culture: Inoculate Achromobacter sp. D-12 on the slant medium and culture at 30°C for 6 days to obtain the slant of the bacteria; the slant medium is prepared according to the following composition: yeast powder 10g, peptone 5.0g, chlorine Sodium chloride 10.0g, agar 15.0g, water 1000ml;

[0054] (2) Seed culture: Pick an inoculation ring from the slant of the bacteria in step (1) and inoculate it into the inorganic salt culture solution, and cultivate it at 30°C for 6 days to obtain the seed solution; the final concentration of the inorganic salt culture solution is the same as Embodiment 1;

[0055] (3) Expansion culture: Inoculate the seed solution obtained in step (2) into LB liquid medium (100 mL) at an inoculum volume concentration of 10-20 %, and culture with shaking at 30°C and 150 rpm until the logarithmic growth phase, and obtain Bacterial solution, centrifuge the bacter...

Embodiment 3

[0057] Embodiment 3: Acetochlor degradation experiment

[0058] 1) Detection of bacterial concentration and acetochlor content in inorganic salt culture solution:

[0059] The growth of bacteria is detected by ultraviolet spectrophotometer, which is expressed by measuring the absorbance value of bacteria in the culture solution at 600nm.

[0060] In this experiment, reversed-phase high-performance liquid chromatography was used to detect the residual amount of acetochlor in the inorganic salt culture medium. Reverse-phase high-performance liquid chromatography detection conditions: mobile phase is acetonitrile: water = 75:25 (volume ratio), analytical column is Waters C18 column (4.6×250mm, 5μm), flow rate is 1ml / min, injection volume is 20μl , the column temperature is 30°C.

[0061] 2) Acetochlor degradation experiment:

[0062] Take four 250ml Erlenmeyer flasks, add 100ml of inorganic salt culture solution to each, add acetochlor after high-pressure steam sterilization (...

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Abstract

The invention provides novel achromobacter sp. D-12 for efficiently degrading acetochlor of amide herbicides and application of the novel achromobacter sp. D-12. The strain was preserved in the China Center for Type Culture Collection (CCTCC) in Wuhan University, Wuhan, China on September, 27th, 2012, and the preservation number is CCTCC No: M2012386. The achromobacter sp. D-12 can be applied to the degradation of the acetochlor in a water body in a direct adding mode, and can be used for safely, efficiently and quickly degrading the acetochlor remained in the water; and a microbial inoculum containing the strain is easy to prepare, low in cost and convenient to use, and has a good application prospect.

Description

(1) Technical field [0001] The invention relates to a novel strain of Achromobacter sp. D-12 which efficiently degrades the amide herbicide acetochlor and its application. (2) Background technology [0002] Acetochlor is a widely used herbicide, mainly used for weeding corn, soybean, peanut, cotton, potato and other crops. It was successfully developed by Monsanto in 1971. It is currently the most widely used herbicide in the world. It is one of the varieties of herbicides and also one of the most widely used herbicides in my country. The molecular formula of acetochlor is C 14 h 20 ClNO 2 , the structural formula is as follows. [0003] [0004] Because acetochlor has medium to high water solubility and relatively low soil adsorption constant, the amide herbicides applied to farmland are easy to transfer to shallow groundwater through infiltration or enter surface water with rainwater runoff, which is harmful to the water body environment. create pollution. Toxicolo...

Claims

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Application Information

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IPC IPC(8): C12N1/20A62D3/02C02F3/34C12R1/025A62D101/04A62D101/28C02F101/38
Inventor 徐超丁静茴
Owner ZHEJIANG UNIV OF TECH
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